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Nucleic Acids Research, 1988, Vol. 16, No. 14 6411-6426
© 1988

Articles

Replication of single-stranded DNA templates by primase-polymerase complexes of the yeast, Saccharomyces cerevisiae

Esther E. Biswas and Subhasis B. Biswas*

Division of Endocrinology, BRB 10-047, University of Maryland School of Medicine Baltimore, MD 21201, USA

*To whom correspondence should be addressed

Received March 1, 1988. A partially purified primase-polymerase complex from the yeast, Saccharomyces cerevisiae was capable of replicating a single stranded circular phage DNA into a replicative form with high efficiency. The primase-polymerase complex exhibited primase activity and polymerase activity on singly primed circular ssDNA as well as on gapped DNA. In addition, it was able to replicate an unprimed, single-stranded, circular phage DNA through a coupled primase-polymerase action. On Biogel A-0.5m filtration the primase-polymerase activities appeared in the void volume, demonstrating a mass of greater than 500 kilodaltons. Primase and various primase-polymerase complexes synthesized unique primers on single stranded DNA templates and the size distribution of primers was dependent on the structure of the DNA and the nature of the primase-polymerase assembly.


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